The study of regulatory effects of Pdx-1, MafA and NeuroD1 on the activity of porcine insulin promoter and the expression of human islet amyloid polypeptide

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• 作者：Xiao-Dan Liu (1)
  Jin-Xue Ruan (1)
  Ji-Han Xia (1)
  Shu-Lin Yang (1)
  Jun-Hua Fan (1)
  Kui Li (1)
  
• 关键词：Insulin promoter ; Transcription factor ; Luciferase ; Human islet amyloid polypeptide ; PK15 cell
• 刊名：Molecular and Cellular Biochemistry
• 出版年：2014
• 出版时间：September 2014
• 年：2014
• 卷：394
• 期：1-2
• 页码：59-66
• 全文大小：450 KB
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• 作者单位：Xiao-Dan Liu (1)
  Jin-Xue Ruan (1)
  Ji-Han Xia (1)
  Shu-Lin Yang (1)
  Jun-Hua Fan (1)
  Kui Li (1)
  
  1. Key Laboratory for Farm Animal Genetic Resources and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing, 100193, People鈥檚 Republic of China
  
• ISSN：1573-4919

文摘

The purpose of the present study was to determine the activation of porcine insulin promoter (PIP) by three transcription factors: pancreatic and duodenal homeobox 1 (Pdx-1), v-maf musculoaponeurotic fibrosarcoma oncogene (MafA) and neurogenic differentiation 1 (NeuroD1) in non-beta islet cells cultured in vitro. In addition, the expression of the exogenous human islet amyloid polypeptide (hIAPP) gene driving by PIP in porcine kidney 15 (PK15) cells co-transfected with these transcription factors was also examined. In the present study, a series of vectors for gene overexpression were constructed, including pGL3-Pdx-1, pGL3-MafA, pGL3-NeuroD1, pGL3-PIP-LUC and pcDNA3.1-PIP-hIAPP. The dual-luciferase reporter assay showed that the PIP activity was increased in PK15 cells when overexpressing the exogenous transcription factors Pdx-1, MafA and NeuroD1. Introducing the PIP-hIAPP expression vector into PK15 cells combined with exogenous Pdx-1, MafA and NeuroD1 resulted in the efficient expression of hIAPP at the gene level, but not the protein. The current systematic porcine insulin promoter analysis provided the basic information for future utilization of porcine insulin.